Abstract:
:Neurofilaments were assembled in vitro from the high speed supernatant of mammalian CNS homogenate in a disassembly buffer containing 4-morpholine-ethane sulfonic acid, MgCl2 and EGTA at 4 degrees C in the presence of 4 M glycerol. The assembled neurofilaments were depolymerized by dialysis against the disassembly buffer and repolymerized by the addition of glycerol to the clarified supernatant obtained afer disassembly. The filament assembly reaction was complete in less than 30 s as measured by turbidimetric changes at 415 nm and did not require any added nucleotide. No assembly of filaments was detected when using frozen tissue. The assembled filaments corresponded to the enrichment of neurofilament triplet, the 210,000, 160,000 and 70,000 dalton polypeptides on SDS-polyacrylamide gels and appeared morphologically and immunochemically identical to neurofilaments isolated by axonal flotation methods. These studies demonstrate in vitro assembly of neurofilaments under native conditions which raises the possibility that like microtubules, neurofilaments or a subpopulation of neurofilaments might be in a dynamic state of assembly--disassembly in situ.
journal_name
Brain Resjournal_title
Brain researchauthors
Iqbal K,Grundke-Iqbal I,Merz PA,Wisniewski HM,Zaidi Tdoi
10.1016/s0006-8993(87)80010-0subject
Has Abstractpub_date
1987-07-01 00:00:00pages
163-72issue
2eissn
0006-8993issn
1872-6240pii
S0006-8993(87)80010-0journal_volume
388pub_type
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